Non-telecentric two-photon microscopy for 3D random access mesoscale imaging

F. K. Janiak, P. Bartel, M. R. Bale, T. Yoshimatsu, E. Komulainen, M. Zhou, K. Staras, L. L. Prieto-Godino, T. Euler, M. Maravall, T. Baden

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4 Scopus citations


Diffraction-limited two-photon microscopy permits minimally invasive optical monitoring of neuronal activity. However, most conventional two-photon microscopes impose significant constraints on the size of the imaging field-of-view and the specific shape of the effective excitation volume, thus limiting the scope of biological questions that can be addressed and the information obtainable. Here, employing a non-telecentric optical design, we present a low-cost, easily implemented and flexible solution to address these limitations, offering a several-fold expanded three-dimensional field of view. Moreover, rapid laser-focus control via an electrically tunable lens allows near-simultaneous imaging of remote regions separated in three dimensions and permits the bending of imaging planes to follow natural curvatures in biological structures. Crucially, our core design is readily implemented (and reversed) within a matter of hours, making it highly suitable as a base platform for further development. We demonstrate the application of our system for imaging neuronal activity in a variety of examples in zebrafish, mice and fruit flies.

Original languageEnglish
Article number544
JournalNature communications
Issue number1
StatePublished - Dec 2022


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